Histone H2B, acetylated, Human, BioAssay(TM) ELISA Kit

The Acetylated Histone H2B Sandwich ELISA Kit is a solid phase sandwich enzyme- linked immunosorbent assay (ELISA) that detects endogenous levels of acetylated lysines on Histone H2B. The Histone H2B Antibody has been coated onto the microwells. After incubation with cell lysates, Histone H2B is captured by the coated antibody. Following extensive washing, Acetylated-Lysine Mouse Mab is added to detect the acetylated lysines on the Histone H2B protein. Anti-mouse IgG,HRP linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of acetylated Histone H2B. Antibodies in this kit are custom formulations specific to this kit. Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of four core histone proteins (H2A, H2B, H3 and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various post-translational modifications, including acetylation, phosphorylation, methylation and ubiquitination (2-5). These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, on gene expression (6). In most species, histone H2B is primarily acetylated at lysines 5, 12, 15 and 20 (4,7). Histone H3 is primarily acetylated at lysines 9, 14, 18 and 23 (2,3). Acetylation at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms (2,3). Phosphorylation at Ser10, Ser28 and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8,9,10). Phosphorylation of Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation of H3 Thr3 in prophase and its dephosphorylation during anaphase (11).